text: In general, the recovery of fluorescence will not be dominated by either simple isotropic diffusion, or by a single simple unbinding rate. There will be both diffusion and binding, and indeed the diffusion constant may not be uniform in space, and there may be more than one type of binding sites, and these sites may also have a non-uniform distribution in space. Flow processes may also be important. This more complex behavior implies that a model with several parameters is required to describe the data; models with only either a single diffusion constant D or a single off rate constant, k, are inadequate.
  There are models with both diffusion and reaction. Unfortunately, a single FRAP curve may provide insufficient evidence to reliably and uniquely fit (possibly noisy) experimental data. Sadegh Zadeh  et al.  have shown that FRAP curves can be fitted by different pairs of values of the diffusion constant and the on-rate constant, or, in other words, that fits to the FRAP are not unique. This is in three-parameter (on-rate constant, off-rate constant and diffusion constant) fits. Fits that are not unique, are not generally useful.
  Thus for models with a number of parameters, a single FRAP experiment may be insufficient to estimate all the model parameters. Then more data is required, e.g., by bleaching areas of different sizes, determining some model parameters independently, etc.